Amine compounds and inhibiting neurotransmitter reuptake

ABSTRACT

The invention relates to amine compounds as well as methods and materials involved in modulating neurotransmitter reuptake. Specifically, the invention provides amine compounds, methods for synthesizing amine compounds, and methods for inhibiting neurotransmitter reuptake.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.11/694,656, filed Mar. 30, 2007; which is a divisional of U.S.application Ser. No. 11/137,313, filed May 25, 2005; which is adivisional of U.S. application Ser. No. 10/755,893, filed Jan. 12, 2004(now U.S. Pat. No. 6,914,080); which is a divisional of U.S. applicationSer. No. 09/907,377, filed Jul. 17, 2001 (now U.S. Pat. No. 6,700,018).The disclosure of the prior applications are considered part of (and areincorporated by reference in) the disclosure of this application.

BACKGROUND

1. Technical Field

The invention relates to amine compounds as well as methods andmaterials involved in modulating neurotransmitter reuptake.

2. Background Information

Neuronal signals are transmitted from cell to cell at specialized sitesof contact known as synapses. The usual mechanism of transmission isindirect. The cells are electrically isolated from one another, thepresynaptic cell being separated from the postsynaptic cell by a narrowsynaptic cleft. A change of electrical potential in the presynaptic celltriggers it to release signaling molecules known as neurotransmitters.The neurotransmitters rapidly diffuse across the synaptic cleft andprovoke an electrical change in the postsynaptic cell by binding toneurotransmitter-gated ion channels. After release, the excessneurotransmitters are rapidly removed, either by specific enzymes in thesynaptic cleft or by reuptake into the presynaptic cell or surroundingglial cells. Reuptake is mediated by a variety of neurotransmittertransporters. Rapid removal ensures both spatial and temporal precisionof signaling at a synapse. For example, rapid reuptake can preventexcess neurotransmitters from influencing neighboring cells and canclear the synaptic cleft before the next pulse of neurotransmitterrelease so that the timing of repeated, rapid signaling events isaccurately communicated to the postsynaptic cell.

An imbalance of neurotransmitters in the brain can occur when not enoughneurotransmitter is made and released from presynaptic cells or thereuptake of neurotransmitters by presynaptic cells is too rapid. Ifneurotransmitters such as serotonin, norepinephrine, or dopamine are notmade and released in effective amounts or are cleared from the synapticcleft too quickly, then cell-to-cell communication can be affected.Clinical manifestations of such imbalances include depression andrelated anxiety disorders. Serotonin-, norepinephrine-,dopamine-reuptake inhibitors (SNDRIs) represent a class of potent,wide-spectrum antidepressant medications that inhibit the reuptake ofthese neurotransmitters back into presynaptic cells. Inhibitingneurotransmitter reuptake can increase the amount of neurotransmitterpresent in the synapse, thus helping to normalize the transmission ofneuronal signals and alleviate the symptoms of depression and relatedanxiety disorders.

SUMMARY

The invention relates to amine compounds as well as methods andmaterials involved in modulating neurotransmitter reuptake.Specifically, the invention provides amine compounds, methods forsynthesizing amine compounds, and methods for inhibitingneurotransmitter reuptake. The amine compounds provided herein can beused as potent, wide-spectrum antidepressant medications for inhibitingneurotransmitter reuptake and treating anxiety disorders. In addition,the methods provided herein for synthesizing amine compounds allow forsynthesis in a reliable and efficient manner.

In general, the invention features a composition containingN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. TheN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can contain(2R,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine or(2S,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. TheN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can contain(2R,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine or(2S,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. TheN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can contain (a)two compounds selected from the following group:(2R,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2S,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2R,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, and(2S,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine; (b)three compounds selected from the following group:(2R,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2S,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2R,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, and(2S,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine; or (c)(2R,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2S,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2R,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, and(2S,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. Thecomposition can contain a pharmaceutically acceptable carrier. At leastabout 35 percent of the composition (e.g., at least about 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, or 99 percent of the composition)can be the N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

In another embodiment, the invention features a composition containingN-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine. TheN-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine can contain(2R,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine or(2S,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine. TheN-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine can contain(2R,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine or(2S,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine. TheN-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine can contain(a) two of the compounds selected from the following group:(2R,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine; (b)three of the compounds selected from the following group:(2R,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine; or(c) (2R,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine. Thecomposition can contain a pharmaceutically acceptable carrier. At leastabout 35 percent of the composition (e.g., at least about 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, or 99 percent of the composition)can be the N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine.

Another embodiment of the invention features a composition containing3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. The3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain(2R,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine or(2S,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. The3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain(2R,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine or(2S,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. The3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain (a) twocompounds selected from the following group:(2R,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine; (b) threecompounds selected from the following group:(2R,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine; or (c)(2R,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. The compositioncan contain a pharmaceutically acceptable carrier. At least about 35percent of the composition (e.g., at least about 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, or 99 percent of the composition) can be the3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine.

Another embodiment of the invention features a composition containingN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. TheN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain(2R,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine or(2S,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. TheN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain(2R,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine or(2S,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. TheN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain(a) two compounds selected from the following group:(2R,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine; (b)three compounds selected from the following group:(2R,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine; or(c) (2R,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. Thecomposition can contain a pharmaceutically acceptable carrier. At leastabout 35 percent of the composition (e.g., at least about 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, or 99 percent of the composition)can be the N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine.

Another embodiment of the invention features a composition containingN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. TheN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain(2R,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine or(2S,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. TheN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain(2R,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine or(2S,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. TheN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine can contain (a)two compounds selected from the following group:(2R,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl) pentylamine,(2S,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine; (b)three compounds selected from the following group:(2R,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine; or (c)(2R,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2S,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, and(2S,3R)-N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. Thecomposition can contain a pharmaceutically acceptable carrier. At leastabout 35 percent of the composition (e.g., at least about 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, or 99 percent of the composition)can be the N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine.

Another embodiment of the invention features a composition containing3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine with the3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine being(2R,3S)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine or(2S,3R)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. The3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can be(2R,3S)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. The3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can be(2S,3R)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. The compositioncan lack (2R,3R)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine and(2S,3S)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. The compositioncan contain a pharmaceutically acceptable carrier. At least about 35percent of the composition (e.g., at least about 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95, or 99 percent of the composition) can be the3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

Another embodiment of the invention features a composition containingN,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine with theN,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine being(2R,3S)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine or(2S,3R)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. TheN,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can be(2R,3S)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. TheN,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can be(2S,3R)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. Thecomposition can lack(2R,3R)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine and(2S,3S)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. Thecomposition can contain a pharmaceutically acceptable carrier. At leastabout 35 percent of the composition (e.g., at least about 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, or 99 percent of the composition)can be the N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

In another aspect, the invention features a method for inhibitingneurotransmitter reuptake in a mammal (e.g., human). The method includesadministering a composition containing at least one compound to themammal, wherein the composition contains at least one compound selectedfrom the following group:N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,(2R,3S)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2S,3R)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2R,3S)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, and(2S,3R)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. Theneurotransmitter reuptake can be norepinephrine or epinephrine reuptake.The neurotransmitter reuptake can be dopamine reuptake or serotoninreuptake. At least about 35 percent of the composition (e.g., at leastabout 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 99 percent ofthe composition) can be the compound.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention pertains. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and not intendedto be limiting.

Other features and advantages of the invention will be apparent from thefollowing detailed description, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. 1 is a diagram ofN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine and its fourstereoisomers.

FIG. 2 is a diagram of N-methyl-3-hydroxy-4,4-dimethyl-2-(2′ naphthyl)pentylamine and its four stereoisomers.

FIG. 3 is a diagram of 3-hydroxy-4-methyl-2-(2′-naphthyl) pentylamineand its four stereoisomers.

FIG. 4 is a diagram of N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine and its four stereoisomers.

FIG. 5 is a diagram of N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine and its four stereoisomers.

FIG. 6 is a diagram of(2S,3R)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine and(2R,3S)-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

FIG. 7 is a diagram of(2S,3R)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine and(2R,3S)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

DETAILED DESCRIPTION

The invention relates to amine compounds as well as methods andmaterials involved in modulating neurotransmitter reuptake.Specifically, the invention provides amine compounds such as3-hydroxy-pentylamine and 3-hydroxy-propylamine compounds, methods forsynthesizing amine compounds, and methods for inhibitingneurotransmitter reuptake. For example, the invention providesN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,N-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, andN,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine compounds. Itis understood that a particular 3-hydroxy-pentylamine or3-hydroxy-propylamine compound includes any one of that compound'sstereoisomers as well as any combination thereof. For example, anN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine compound can be(2R,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2S,3s)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2R,3s)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, or(2S,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, or anycombination of(2R,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2S,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,(2R,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, and(2S,3R)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

The invention also provides methods of synthesizing amine compounds suchas 3-hydroxy-pentylamine and 3-hydroxy-propylamine compounds. Forexample, 3-hydroxy-pentylamine and 3-hydroxy-propylamine compounds canbe synthesized by a variety of organic chemistry techniques including,without limitation, carbamate reduction, N,N dimethylation, aldolreduction, and nitrile reduction. An N-methyl secondary amine compoundof the invention can be synthesized by treating a primary amine withethyl chloroformate to produce a carbamate intermediate, and thenreducing the carbamate intermediate with lithium aluminum hydride. Forexample, the carbamate formed by treating3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine with ethyl chloroformatecan be reduced with lithium aluminum hydride to yieldN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. Other examplesof N-methyl secondary amine compounds that can be synthesized in thismanner include, without limitation,N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine,N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, andN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine.

An N,N-dimethyl tertiary amine compound of the invention can besynthesized by reductive methylation of a primary amine. For example,3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine can be treated withformalin, zinc chloride in methanol, and sodium cyanoborohydride toproduce N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.Another example of an N,N-dimethyl tertiary amine compound that can besynthesized in this manner isN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine.

A primary amine compound of the invention can be synthesized by aldolreduction. An aldehyde can be reacted with an appropriate lithiatedarylacetonitrile to produce an aldol, which can then be reduced withaluminum chloride modified lithium aluminum hydride to form a primaryamine. For example, 3-hydroxy-4-methyl-2-(2′-naphthyl)pentanenitrileprepared from isobutyraldehyde and 2-naphthylacetonitrile can be reducedto 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. Another example of aprimary amine compound that can be synthesized in this manner is3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

Any amine compound provided herein can be resolved to a pure enantiomerby classical resolution using enantiomerically pure acids including,without limitation, (+)- and (−)-tartaric acid, (+)- and(−)-ditoluoyl-tartaric acid, and (+)- and (−)-camphorsulfonic acid. Inaddition, any amine compound or enantiomer thereof provided herein canbe chemically converted from its free base form to a pharmaceuticallyacceptable salt by reacting the free base with an equivalent amount ofany acid that forms a non-toxic salt. Such acids can be either inorganicor organic including, without limitation, hydrochloric acid, hydrobromicacid, fumaric acid, maleic acid, succinic acid, sulfuric acid,phosphoric acid, tartaric acid, acetic acid, citric acid, and oxalicacid. Any amine compound or pharmaceutically acceptable salt thereof canbe administered to a mammal in combination with a carrier. Such carriersinclude, without limitation, sterile aqueous or non-aqueous solutions,suspensions, and emulsions. Examples of non-aqueous solvents include,without limitation, propylene glycol, polyethylene glycol, vegetableoils, and injectable organic esters. Aqueous carriers include, withoutlimitation, water, alcohol, saline, and buffered solutions.Preservatives, flavorings, and other additives such as, for example,antimicrobials, anti-oxidants, chelating agents, inert gases, and thelike can also be present. It will be appreciated that any amine compoundprovided herein that is to be administered to a mammal can contain zero,one, or more than one commonly known pharmaceutically acceptablecarriers.

The invention provides methods for using amine compounds such as3-hydroxy-pentylamine and 3-hydroxy-propylamine compounds to inhibitneurotransmitter reuptake in a mammal. The term “inhibit” as used hereinwith respect to neurotransmitter reuptake refers to any reduction inneurotransmitter reuptake. For example, a reduction in neurotransmitterreuptake greater than zero percent (e.g., 0.1, 0.5, 1, 2, 5, 10, 25, 50,75, or 99 percent) is considered an inhibition of neurotransmitterreuptake. Any method can be used to assess whether or notneurotransmitter reuptake has been inhibited in a mammal. Such methodscan be qualitative or quantitative. An example of a qualitative methodincludes assessing whether or not a mammal with depression or a relatedanxiety disorder experiences loss of pleasure in daily activities,significant weight loss or gain, changes in mobility (e.g., lethargy,nervousness), feelings of worthlessness, diminished ability toconcentrate, or suicidal thoughts to a lesser extent following treatmentwith an amine compound provided herein than the extent experiencedbefore treatment. Alternatively, such methods can be quantitative. Forexample, the concentration of serotonin in a platelet sample from amammal after treatment with an amine compound can be measured andcompared to the concentration of serotonin in a platelet sample from thesame mammal before treatment with that amine compound. If theconcentration of serotonin after treatment is reduced compared to theconcentration of serotonin before treatment, then that amine compoundinhibited neurotransmitter reuptake in that mammal.

To inhibit neurotransmitter reuptake, an effective amount of any aminecompound provided herein can be administered to a mammal. The term“effective” as used herein refers to any amount that induces a desiredlevel of neurotransmitter reuptake inhibition while not inducingsignificant toxicity in the mammal. Such an amount can be determinedusing the methods and materials provided herein. An effective amount ofan amine compound or formulation containing an amine compound can be anyamount that reduces, prevents, or eliminates an anxiety disorder uponadministration to a mammal without producing significant toxicity tothat mammal. Some amine compounds may have a relatively broadconcentration range that is effective while others may have a relativelynarrow effective concentration range. In addition, the effective amountcan vary depending upon the specific mammal or anxiety disorder to betreated because certain mammals and anxiety disorders can be more orless responsive to a particular amine compound. Such effective amountscan be determined for individual amine compounds using commonlyavailable or easily ascertainable information involving equilibriumdissociation constants, mammal toxicity concentrations, andbioavailability. For example, non-toxic amine compounds typically can bedirectly or indirectly administered to a mammal in any amount thatreduces, prevents, or eliminates an anxiety disorder in that mammal.Using the information provided herein, such effective amounts can alsobe determined by routine experimentation in vitro or in vivo. Forexample, a patient having an anxiety disorder can receive directadministration of an amine compound in an amount close to theequilibrium dissociation constant (i.e., K_(d)) calculated from in vitroanalysis sufficient to inhibit the uptake of a particularneurotransmitter. If the patient fails to respond, then the amount canbe increased by, for example, two fold. After receiving this higherconcentration, the patient can be monitored for both responsiveness tothe treatment and toxicity symptoms, and adjustments made accordingly.

To help determine effective amounts of different amine compounds, it canbe useful to refer to an effective amount equivalent based on theeffective amount of a common drug used to treat anxiety disorders. Forexample, the direct administration of 0.30 mg/kg Prozac daily for threeweeks to a mammal can be an effective amount for treating anxietydisorders. The effects produced by this effective amount can be used asa reference point to compare the effects observed for other aminecompounds used at varying concentrations. Once an equivalent effect isobserved, then the specific effective amount for that particular aminecompound can be determined. In this case, that particular amount wouldbe termed a Prozac effective amount equivalent.

The ability of an amine compound to inhibit neurotransmitter reuptakealso can be assessed in vitro. For example, the level of serotoninreuptake can be determined by measuring the amount of radiolabeledserotonin taken up by synaptosomes purified from a tissue sourceabundant in serotonin transporters (e.g., rat brain cortical tissue).Rat brain cortical tissue can be isolated to produce neuronal membranefragments such that the membrane fragments close back on themselves toform synaptosomes that retain functional serotonin transporters. Theserotonin transporters concentrate serotonin by transporting it from thefluid in which the synaptosomes are suspended to the interior of thesynaptosomes. If the serotonin in the suspension fluid is radiolabeled,then the level of serotonin reuptake can be measured by counting theradioactivity in the synaptosomal pellet obtained by centrifugation. Theability of an amine compound to inhibit the level of serotonin reuptakecan be determined by adding different concentrations to aliquots of thesame synaptosomal preparation. For example, the potency ofN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine as aninhibitor of serotonin reuptake can be measured by (1) adding differentconcentrations ofN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine to aliquotsof synaptosomes purified from rat brain cortical tissue, (2) adding thesame concentration of radiolabeled serotonin to each aliquot, (3)allowing the serotonin transporters to concentrate the radiolabeledserotonin in the synaptosomes, and (4) counting the radioactivity in thesynaptosomal pellet of each aliquot obtained after centrifugation. Aminecompounds with a higher potency will more effectively inhibit reuptakethus resulting in less detectable radioactivity in the synaptosomalpellet.

In another in vitro example, intact cultured mammalian cells expressinga particular recombinant neurotransmitter transporter can be used toassess the ability of an amine compound to inhibit neurotransmitterreuptake. For example, the potency ofN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine as an inhibitorof norepinephrine transport can be measured using cultured mammaliancells expressing the norepinephrine transporter. In addition, thepotency of a particular amine compound to inhibit multipleneurotransmitter transporters can be measured. For example, the potencyof N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine as aninhibitor of both serotonin and norepinephrine transport can be measuredusing separate cultured mammalian cells expressing the serotonintransporter and cultured mammalian cells expressing the norepinephrinetransporter. It is understood that measured neurotransmitter transportlevels are compared to controls. Controls include, without limitation,vehicle only as well as known inhibitors such as Prozac, Paxil, Effexor,or Serzone.

In addition, the potency of an amine compound to inhibit the reuptake ofdifferent neurotransmitters can be assessed by determining theequilibrium dissociation constant (i.e., K_(d)) of that particular aminecompound for a particular neurotransmitter transporter. Typically, theK_(d) value is determined as described elsewhere (Tatsumi et al, Eur. J.Pharmacol., 340:249-258 (1997)). Once determined, the K_(d) value for aparticular amine compound can be used to compare that compound's potencywith the potency of other amine compounds or other known inhibitors. Forexample, if a particular compound has a K_(d) of 0.3 nM for theserotonin transporter and a K_(d) of 14.6 nM for the norepinephrinetransporter, then that particular compound can be characterized ashaving a greater ability to inhibit serotonin reuptake compared tonorepinephrine reuptake. Likewise, if a first amine compound has a K_(d)of 0.3 nM for the serotonin transporter and a second amine compound hasa K_(d) of 6.3 nM for the serotonin transporter, then the first aminecompound can be characterized as having a greater ability to inhibitserotonin reuptake compared to the second amine compound.

Various factors can influence the actual effective amount used for aparticular application. For example, the frequency of administration,duration of treatment, rate of metabolism of the drug, combination ofother amine compounds, and site of administration may require anincrease or decrease in the actual effective amount administered.

The frequency of administration can be any frequency that reduces,prevents, or eliminates an anxiety disorder or depression in a mammalwithout producing significant toxicity to the mammal. For example, thefrequency of administration can be from about once a day to about once amonth, or more specifically, from about twice a day to about once aweek. In addition, the frequency of administration can remain constantor can be variable during the duration of treatment. As with theeffective amount, various factors can influence the actual frequency ofadministration used for a particular application. For example, theeffective amount, duration of treatment, rate of metabolism of the drug,combination of other amine compounds, and site of administration mayrequire an increase or decrease in administration frequency.

An effective duration for amine compound administration can be anyduration that reduces, prevents, or eliminates an anxiety disorder in amammal without producing significant toxicity to the mammal. Thus, theeffective duration can vary from several days to several weeks, months,or years. In general, the effective duration for the treatment of ananxiety disorder can range in duration from several days to severalyears. Once the amine compound administrations are stopped, however, thetreated anxiety disorder may return. Thus, the effective duration forthe prevention of an anxiety disorder can last in some cases for as longas the individual is alive.

Multiple factors can influence the actual effective duration used for aparticular treatment or prevention regimen. For example, an effectiveduration can vary with the frequency of amine compound administration,effective amine compound amount, combination of multiple aminecompounds, and site of administration. It is noted that diagnosticalgorithm methods can be devised to determine or reflect appropriateeffective doses, durations, and frequencies.

The level of toxicity, if any, can be determined by assessing a mammal'sclinical signs and symptoms before and after administering a knownamount of a particular composition. It is noted that the effectiveamount of a particular composition administered to a mammal can beadjusted according to a desired outcome as well as the mammal's responseand level of toxicity. Significant toxicity can vary for each particularmammal and each particular composition.

Any combination of amine compounds can be administered to a mammal. Forexample, two amine compounds can be administered together to a mammal toinhibit norepinephrine reuptake in that mammal. In another example, oneor more compounds that can inhibit serotonin reuptake and one or morecompounds that can inhibit dopamine reuptake can be administeredtogether to a mammal to inhibit both serotonin and dopamine reuptake inthat mammal. The efficacy of such combinations can be assessed using themethods and materials provided herein.

An amine compound or combination of amine compounds can be administeredto any part of a mammal's body. For example, an amine compound can bedelivered to, without limitation, spinal fluid, blood, lungs,intestines, muscle tissues, skin, joints, peritoneal cavity, or brain ofa mammal. In addition, an amine compound or combination of aminecompounds can be administered intravenously, intraperitoneally,intramuscularly, subcutaneously, intrathecally,intracerebroventricularly, or intradermally, orally, by inhalation, orby gradual perfusion over time. The duration of treatment can be anylength of time from as short as one day to as long as the life span ofthe mammal (e.g., many years). For example, an amine compound can beadministered daily for three months or ten years. It is also noted thatthe frequency of treatment can be variable. For example, an aminecompound can be administered once (or twice, three times, etc.) daily,weekly, monthly, or yearly.

The invention will be further described in the following examples, whichdo not limit the scope of the invention described in the claims.

EXAMPLES Example 1 Human Transporter Binding Studies

Human embryonic kidney (HEK-293) cells stably transfected andconstitutively expressing the human norepinephrine transporter (hNET;Pacholczyk et al., Nature, 350:350-354 (1991)), the human dopaminetransporter (hDAT; Pristupa et al., Mol. Pharmacol., 45:125-135 (1994)),or the human serotonin transporter (hSERT; Ramamoorthy et al., Proc.Natl. Acad. Sci. U.S.A. 90:2542-2546 (1993)) were grown and passaged in150-mm petri dishes with 17.5 ml of Dulbecco's modified Eagle's medium(MEM; Mediatech Inc., Herndon, Va.) containing 0.1 mM non-essentialamino acid solution for MEM (Mediatech Inc.), 5% (v/v) fetal clonebovine serum product (Hyclone Laboratories, Logan, Utah), and 1 U/μLpenicillin/streptomycin solution (Mediatech Inc.). The cells wereincubated in 10% CO₂, 90% air at 37° C. and 100% humidity. The hNET cellcultures contained 250 μg/mL geneticin sulfate. The cells were grown to70-80% confluency prior to harvesting.

Cell membranes containing hSERT, hNET, or hDAT were prepared from thecell lines to assay ligand binding for each of the transporters.Briefly, the cell medium was removed by aspiration, and the cells werewashed with 4 mL modified Puck's D1 solution (solution 1; Richelson etal. in “Methods in Neurotransmitter Receptor Analysis” Yamamura, H. I.;Enna, S. J.; Kuhar, M. J. Eds.; New York, Raven Press, 1990, pp147-175). The washed cells were incubated for 5 minutes at 37° C. in 10mL solution 1 containing 100 mM ethylene glycol-bisN,N,N′,N′-tetraacetic acid (EGTA). The cells were then scraped from theflask surface with a rubber spatula, placed into a centrifuge tube, andcollected by centrifugation at 1000×g for 5 minutes at 4° C. Theresulting supernatant was discarded, and the cell pellet was resuspendedin 0.5 to 1.0 mL of the appropriate binding buffer (described below).The resuspended cell pellet was homogenized using a Polytron for 10seconds at setting 6. The resulting homogenate was centrifuged at about36,000×g for 10 minutes at 4° C. The supernatant was discarded, and thepellet was resuspended in the same volume of the appropriate bindingbuffer and centrifuged again. The supernatant was discarded, and thefinal pellet containing cell membranes was resuspended in theappropriate binding buffer and stored at −80° C. until use. The finalprotein concentration was determined by the Lowry assay using bovineserum albumin as a standard (Lowry et al., J. Biol. Chem. 193:265-275(1951)).

Radioligand binding assays for the indicated transporters were performedas follows. To assess binding to the cloned hSERT, cells expressinghSERT were homogenized in 50 mM Tris-HCl with 120 mM NaCl and 5 mM KCl(pH 7.4). The binding reaction consisted of 30 μg cell membrane protein,1.0 nM [³H]imipramine (imipramine hydrochloride, benzene ring-3H,specific activity 46.5 Ci/mmol; Dupont New England Nuclear, Boston,Mass.), and varying concentrations of either unlabeled imipramine or thetest amine compound. A reaction to determine non-specific bindingconsisted of 15 μg cell membrane protein, 1.0 nM [³H]imipramine, and 1μM final concentration of unlabeled imipramine. The reactions wereincubated at 22° C. for 60 minutes. Following incubation, the reactionswere terminated by rapid filtration through separate GF/B filter stripspretreated with 0.2% polyethylenimine in a 48-well Brandel cellharvester. The cell membrane-containing filter strips were then rinsedfive times with ice-cold 0.9% NaCl. After rinsing, individual filterswere cut from the strip and placed in a scintillation vial containing6.5 mL of Redi-Safe (Beckman Instruments, Fullerton, Calif.).Radioactivity was measured with a Beckman liquid scintillation counter(LS 5000TD).

To assess binding to the cloned hNET, cells expressing hNET werehomogenized in 50 mM Tris-HCl with 300 mM NaCl and 5 mM KCl (pH 7.4).The binding reaction consisted of 25 μg cell membrane protein, 0.5 nM[³H]nisoxetine (nisoxetine HCl, [N-methyl-3H], specific activity 85.0Ci/mmol; Amersham, Arlington Hts., Ill.), and varying concentrations ofeither unlabeled nisoxetine or the test amine compound. A reaction todetermine non-specific binding consisted of 25 μg cell membrane protein,0.5 nM [³H]nisoxetine, and 1 μM final concentration of unlabelednisoxetine. The reactions were incubated at 22° C. for 60 minutes.Following incubation, the reactions were terminated by rapid filtrationthrough separate GF/B filter strips pretreated with 0.2%polyethylenimine in a 48-well Brandel cell harvester. The cellmembrane-containing filter strips were then rinsed five times withice-cold 0.9% NaCl. After rinsing, individual filters were cut from thestrip and placed in a scintillation vial containing 6.5 mL of Redi-Safe(Beckman Instruments, Fullerton, Calif.). Radioactivity was measuredwith a Beckman liquid scintillation counter (LS 5000TD).

To assess binding to the cloned hDAT, cells expressing hDAT werehomogenized in 50 mM Tris-HCl with 120 mM NaCl (pH 7.4). The bindingreaction contained 30 μg cell membrane protein, 1 nM [³H]WIN35428(WIN35428, [N-methyl-3H], specific activity 83.5 Ci/mmol; Dupont NewEngland Nuclear, Boston, Mass.), and varying concentrations of eitherunlabeled WIN35428 or the test amine compound. A reaction to determinenon-specific binding contained 30 μg cell membrane protein, 1 nM[³H]WIN35428, and 10 μM final concentration of unlabeled WIN35428. Thereactions were incubated at 22° C. for 1 hour. Following incubation, thereactions were terminated by rapid filtration through separate GF/Bfilter strips pretreated with 0.2% polyethylenimine in a 48-well Brandelcell harvester. The cell membrane-containing filter strips were thenrinsed five times with ice-cold 0.9% NaCl. After rinsing, individualfilters were cut from the strip and placed in a scintillation vialcontaining 6.5 mL of Redi-Safe (Beckman Instruments, Fullerton, Calif.).Radioactivity was measured with a Beckman liquid scintillation counter(LS 5000TD).

Following the radioligand binding assays, the data were analyzed usingthe LIGAND program (Munson and Rodbard, Analyt. Biochem., 107:220-239(1980)) to provide values for the equilibrium dissociation constants(K_(d)). The program was modified to calculate the Hill coefficient(nH). Data are presented as geometric mean ±S.E.M. of at least threeindependent experiments. One-component models and two-component modelswere compared using the root mean square error of each fit and the Ftest. A low K_(d) for a compound indicates strong binding to thetransporter (i.e., reuptake inhibition).

The compounds exhibited K_(d) values ranging from strong to weaktransporter binding. Compound A exhibited the strongest binding to hNETof all the compounds tested. In addition, compound A exhibited greaterspecificity for hNET than for hSERT or hDAT. Compound B showed greaterspecificity for hDAT than the parent analogue venlfaxine. Compound C wasa more potent inhibitor of hSERT than of hNET or hDAT. Compound C alsoexhibited greater specificity for all three transporters compared tovenlafaxine, indicating that it is a more potent transporter inhibitor.In addition, compounds C1 and C2 showed greater specificity for hSERTthan for hNET of hDAT, and greater specificity for all threetransporters compared to venlafaxine. Compound D exhibited strongerbinding to hSERT than to hNET or hDAT, and was a more potent inhibitorof all three transporters compared to vanlafaxine. Compound D1 showedsimilar specificities for hNET and hSERT, which were greater than thatfor hDAT, indicating that it was a more potent inhibitor of hNET andhSERT compared to hDAT. Compound D2 exhibited greater specificity forhSERT than for hNET or hDAT. Both compounds D1 and D2 showed greaterspecificity for all three transporters compared to venlafaxine.Compounds E and F exhibited greater specificity for hDAT compared tovanlafaxine. These data demonstrated that in vitro human transporterbinding studies can be used to determine the ability of an aminecompound to inhibit neurotransmitter transport. These data alsodemonstrated that amine compounds inhibit neurotransmitter transport invarying degrees at different transporters.

TABLE 1 Human transporter binding data hNET K_(d) hSERT K₆ hDAT K_(d)Compound (nM) SEM (nM) SEM (nM) SEM Nisoxetine 2.4 0.1 Imipramine 1.70.1 WIN35428 29 1 Desipramine 0.83 0.05 17.6 0.7 3190 40 Paroxetine 40 20.13 0.01 490 20 Sertraline 420 20 0.29 0.01 25 2 Venlafaxine 1060 408.9 0.3 9300 50 A 0.25 0.03 6.3 0.3 130 10 B 390 10 C 329 2 5.7 0.5 88080 C1 150 20 7 1 1280 50 C2 140 10 15.1 0.9 1600 200 D 14.6 0.7 0.300.04 180 20 D1 7.4 0.4 4.2 0.3 150 10 D2 60 6 2.6 0.4 800 150 E 420 10A: racemic mixture of (2R,3R)- and(2S,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. B:racemic mixture of (2S,3R)- and(2R,3S)-N-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine. C:racemic mixture of (2R,3S)- and(2S,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. C1:(2R,3S)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. C2:(2S,3R)-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. D: racemicmixture of (2R,3S)-and(2S,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′naphthyl)pentylamine. D1:(2R,3S)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. D2:(2S,3R)-N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. E:racemic mixture of (2S,3R)- and(2R,3S)-3-hydroxy-2-(2′naphthyl)-3-phenylpropylamine. F: racemic mixtureof (2S,3R)- and(2R,3S)-N,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.

Example 2 Neurotransmitter Reuptake in Rat Brain Synaptosomes

Cortical, striatal, and hippocampal tissues are dissected from freshlydecapitated male Sprague-Dawley rats (125-250 g; Harlan Sprague-Dawley,Indianapolis, Ind., USA). The dissected tissues are separatelyhomogenized in 20 volumes of ice-cold 0.32 M sucrose containing 11 mMglucose (pH 7.4) in a glass Potter-Elvehjem homogenizer with Teflon®pestle (8 strokes, 900 rpm). The homogenates are centrifuged at 1,000×gfor 10 minutes. The resulting supernatant is decanted and furthercentrifuged at 20,000×g for 20 minutes. The supernatant is discarded,and the synaptosomes contained in the pellet are gently resuspended inoxygenated incubation buffer containing 10 mM glucose, 20 mM HEPES, 145mM NaCl, 4.5 mM KCl, 1.2 mM MgCl₂, and 1.5 mM CaCl₂ (pH 7.4). Thesynaptosomal protein concentration is determined by the Lowry methodusing bovine serum albumin as a standard (Lowry et al., J. Biol. Chem.193:265-275 (1951)).

Synaptosomal protein (1.0-2.5 mg) is suspended 1 mL oxygenatedincubation buffer containing 10 μM pargyline to inhibit monoamineoxidase activity and 0.2 mg/mL sodium ascorbate. Separate reaction tubescontaining 8 nM levo-[ring-2,5,6-³H]norepinephrine ([³H]NE; 43.7Ci/mmol; New England Nuclear, Boston, Mass.), 4 nM5-[1,2-³H(N)]hydroxytryptamine binoxalate ([³H]5-HT; 23.4 Ci/mmol; NewEngland Nuclear), or 2 nM [7,8-³H]dopamine (DA) ([³H]DA; 47 Ci/mmol;Amersham, Arlington Heights, Ill.) are preincubated for 5 minutes at 37°C. in a shaking water bath (80 oscillations/minute) in the presence ofthe test amine compound. After preincubation, the neurotransmitterreuptake reaction is initiated by the addition of synaptosomal protein.The reaction is stopped after 5 minutes by adding 4 mL ice-cold 0.9%(w/v) sodium chloride. The stopped reactions are rapidly filteredthrough a Whatman GF/B glass fiber filter in a 48-place Brandel cellharvester. The filters containing deposited synaptosomes are then washedwith an additional 8 mL of wash buffer. The washed filters are placed ina scintillation vial containing 5 mL of Redi-Safe (Beckman Instruments,Fullerton, Calif.) and counted. Specific reuptake is calculated as thedifference between the total reuptake (zero unlabelled ligand) andnonspecific reuptake (excess unlabelled ligand).

Example 3 Testing Amine Compounds in Animals and Humans

Test amine compounds are prepared by formulating the amine compoundswith a pharmaceutically acceptable carrier such as saline. To determinethe toxic levels of each amine compound, non-human mammals (e.g., mice,rats, or lower primates) are used. Briefly, 100 rats are randomlyseparated into five groups of 20. One group, group A, receives aplacebo, while the other groups receive a particular dose of a compound,e.g., group B receives the compound at 1 μg/kg, group C receives thecompound at 100 μg/kg, group D receives the compound at 10 mg/kg, andgroup E receives the compound at 100 mg/kg. The rats in each group aredosed according to a prescribed plan for a predetermined period of time,and toxic levels of the administered compound are determined bymeasuring survival or other clinical signs such as aggressiveness,irregular bleeding, and appetite. After determining the toxic level ofan amine compound, tolerability and pharmacokinetic studies areperformed in normal humans.

Once the toxicity, tolerability, and pharmacokinetic studies have beenperformed for an amine compound, 100 human subjects having an anxietydisorder are selected for clinical trials. The subjects are separatedinto two groups of 50. One group, group A, receives a placebo, while theother group, group B, receives the highest possible dose of a compoundthat has been determined not to be toxic to a mammal. The subjects ineach group are dosed according to a prescribed plan for a predeterminedperiod of time, and the ability of the administered amine compound toalleviate anxiety disorder symptoms are determined using standardclinical methods that are used to assess anxiety disorders.

Example 4 Synthesis ofN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, HCl saltSynthesis of Carbamate A

3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine (1.32 g, 4.76 mmol) wasdissolved in CH₂Cl₂ (20 ml), and ethyl chloroformate (550 μL, 5.75 mmol)was added. The resulting milky suspension was stirred for 4 hours atroom temperature. After stirring, water (130 mL) was added to aid in theseparation of the aqueous and organic phases. The aqueous phase wasextracted 4 times with 100 mL ethyl acetate, and the organic phases werecombined, dried over K₂CO₃, filtered, and concentrated in vacuo to yieldcarbamate A (1.65 g, 99%). ¹H NMR (CDCl₃) analysis: δ1.20 (t, J=7.1 Hz,3H), 3.12 (s, 1H), 3.30-3.32 (m, 1H), 3.71-3.74 (m, 1H), 3.86-3.88 (m,1H) 4.08-4.11 (q, J=14.2 Hz, 2H), 4.87 (s, br, 1H), 5.03 (d, J=8 Hz),7.15-7.2 (m, 6H), 7.4-7.48 (m, 2H), 7.52 (s, 1H), 7.1-7.8 (m, 3H). ¹³CNMR (CDCl₃) analysis: δ14.48, 42.88, 53.56, 60.92, ˜76.4, 125.58,125.94, 126.41, 126.61, 127.29, 127.40, 127.46, 127.58, 128.05, 132.34,133.23, 137.22, 142.21, 157.15.

Reduction of carbamate A toN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine

A round bottom flask equipped with a stir bar and water condenser wascharged with carbamate A (1.47 g, 4.21 mmol), lithium aluminum hydride(LiAlH₄; 0.39 g, 10.3 mmol), and dry tetrahydrofuran (THF; 150 mL). Themixture was refluxed under N₂ for 10-12 hours, yielding a blue-graysuspension. The reaction was quenched by adding 1 mL water. Afterquenching the reaction, 10% aqueous NaOH was added with stirring toyield a white solid and a clear organic layer. The organic layer wasdecanted, and 10 mL water was added to the solid. After mixing for 15minutes, the white solid was extracted four times with 20 mL diethylether, and the ether extracts were pooled. The pooled extracts weredried over K₂CO₃, filtered, and concentrated in vacuo to yieldN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine free base (1.14g, 3.91 mmol, 92.9%). This free base was converted to the HCl salt bytreatment with concentrated HCl in methanol, followed by concentrationin vacuo. ¹H NMR (CDCl₃) analysis: δ2.72 (s, 3H), 3.24 (s, br, 1H), 3.21(d, J=10.10 Hz, 1H), 5.17 (d, J=9.15 Hz, 1H), 5.45 (s, br, 1H),7.04-7.14 (m, 4H), 7.21-7.22 (2H), 7.40-7.46 (m, 3H), 7.61-7.71 (m, 3H).¹³C NMR (CDCl₃) analysis: δ34.13, 49.10, 54.87, 79.23, 125.89, 126.01,126.22, 127.00, 127.54, 127.70, 127.86, 128.28, 128.40, 128.47, 132.49,133.21, 135.22, 141.67.

Example 5 Synthesis ofN-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine, HCl saltSynthesis of Carbamate B

3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine (207 mg, 0.706 mmol)was treated with ethyl chloroformate (81 μL, 0.85 mmol) andtriethylamine (0.9 mL, 6.46 mmol) as described in Example 3 to yieldcarbamate B (130.5 mg, 56%).

Synthesis of N-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylaminefrom carbamate B

A three-necked round bottom flask equipped with a stir bar and watercondenser was charged with carbamate B (122.3 mg, 0.37 mmol), LiAlH₄ (31mg, 0.82 mmol) and dry THF (10 mL). The mixture was refluxed under N₂for 4 hours. After refluxing, the reaction was quenched by stirring for1 hour in the presence of 20 mL ether and 10 mL 10% H₂SO₄. The acidlayer was separated and washed two times with 5 mL ether. The etherwashed acid layer was then cooled to 0° C. in an ice bath and adjustedto pH 11 with 2N NaOH. The aqueous layer was then extracted four timeswith 10 mL ether, and the extracts pooled. The pooled extracts werewashed with 25 mL brine and dried over K₂CO₃ overnight. After drying,the pooled extracts were filtered and concentrated in vacuo to yieldN-methyl-3-hydroxy-4,4-dimethyl-2-(2′-naphthyl)pentylamine free base (45mg, 45%). This free base was converted to the HCl salt as described inExample 3. ¹H NMR (CD₃OD) analysis: δ 0.92 (s, 9H), 2.65 (s, 3H),3.15-3.19 (dd, 1H), 3.38-3.44 (dd, 1H), 3.54-3.60 (dd, 1H), 3.87-3.88(d, 1H), 7.42-7.48 (m, 3H), 7.83-7.88 (m, 4H). ¹³C NMR (CD₃OD) analysis:δ 27.54, 35.10, 38.35, 46.82, 56.27, 83.89, 127.74, 127.98, 128.33,129.04, 129.50, 129.55, 131.0, 135.10, 135.92, 140.87.

Example 6 Synthesis of 3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine,HCl salt

3-hydroxy-2-(2′-naphthyl)-3-phenylpropionitrile (0.77 g, 2.8 mmol) wasreduced according to the procedure of Carlier et al. (Carlier et al.,Org. Lett., 2: 2443-2445 (2000)) to yield3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine free base (0.76 g, 98%).This free base was converted to the HCl salt as described in Example 3.¹H NMR (CD₃OD) analysis: δ 3.3-3.33 (1H), 3.40-3.48 (m, 2H), 5.08 (d,J=5.30 Hz, 1H), 7.16-7.83 (m, 12H). ¹³C NMR (CD₃OD) analysis: δ 42.56,52.66, 76.83, 127.09, 127.23, 127.70, 127.92, 128.63, 128.75, 128.83,129.12, 129.27, 129.71, 134.47, 134.81, 135.37, 143.25.

Example 7 Synthesis ofN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, HCl saltSynthesis of Carbamate D

3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine (0.26 g, 0.95 mmol) wastreated with ethyl chloroformate as described in Example 3 yieldingcarbamate D (0.22 g, 67%). ¹H NMR (CDCl₃) analysis: δ 1.15 (t, J=7.10Hz, 3H), 2.63 (s, 1H), 3.24-3.42 (m, 2H), 3.67-3.73 (dt, J=7.1, 13.5 Hz,1H), 4.01-4.06 (dd, 7.10, 14.2 Hz), 4.60 (br, s, 1H), 5.0 (d, 5.5 Hz),7.22-7.82 (m, 12H). ¹³C NMR (CDCl₃) analysis: δ 14.62, 42.98, 53.99,61.06, 75.5, 125.89, 126.18, 126.50, 126.88, 127.69, 127.77, 127.86,128.37, 132.79, 133.40, 135.96, 141.86.

Reduction of carbamate D toN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine

Carbamate D (190 mg, 0.63 mmol) was reduced with LiAlH₄ as described inExample 3 yieldingN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine free base (45.9mg, 25%). This free base was converted to the HCl salt as described inExample 3. ¹H NMR (CD₃OD) analysis: δ 2.64 (s, 3H), 3.39-3.42 (dd,J=4.55, 11.90 Hz, 1H), 3.52-3.62 (m, 2H), 5.09 (d, J=5.30 Hz, 1H),7.11-7.83 (m, 12H). ¹³C NMR (CD₃OD) analysis: δ 34.41, 51.65, 52.40,76.83, 127.16, 127.26, 127.70, 127.96, 128.62, 128.72, 128.84, 129.07,129.28, 129.70, 134.48, 134.79, 135.09, 143.0.

Example 8 Synthesis ofN,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine, HCl salt

3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine (0.20 g, 0.73 mmol) wastreated with formalin, ZnCl₂ (253 mg, 1.86 mmol), and NaBH₃CN (140 mg,2.23 mmol) according to the procedure of Carlier et al. (Carlier et al.,Bioorg. Med. Chem. Lett., 8:487-492 (1998)) to yieldN,N-dimethyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine free base(0.179 g, 80%). This free base was converted to the HCl salt asdescribed in Example 3. ¹H NMR (CD₃OD) analysis: δ 2.84 (s, 3H), 2.87(s, 3H), 3.55-3.59 (dd, J=5.0, 13.0 Hz, 1H), 3.67-3.72 (dt, J=5.0, 10.1Hz, 1H), 3.84-3.88 (dd, J=10.0, 13.0 Hz, 1H), 5.08 (d, J=5.0 Hz, 1H),7.07-7.87 (m, 12H). ¹³C NMR (CD₃OD) analysis: δ 43.57, 45.18, 49.84,60.87, 76.99, 127.25, 127.32, 127.70, 128.04, 128.64, 128.73, 128.89,129.04, 129.36, 129.76, 134.50, 134.66, 134.80, 142.77.

Example 9 Synthesis of 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine,HCl salt Synthesis of Aldol E

3-hydroxy-4-methyl-2-(2′-naphthyl)pentanenitrile was prepared from2-naphthylacetonitrile and isobutyraldehyde according to the procedureof Carlier et al., (Carlier et al., J. Org. Chem., 60:7511-7517 (1995))producing aldol E in 58% yield.

Reduction of aldol E to 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine

Aldol E (1.16 g, 4.9 mmol) was reduced to3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine free base (0.80 g, 59%)according to the procedure of Carlier et al., (Carlier et al., J. Org.Chem., 60:7511-7517 (1995)). This free base was converted to the HClsalt as described in Example 3. ¹H-NMR (CD₃OD) analysis: δ 0.866-0.960(m, 6H), 1.281-1.443 (m, 1H), 3.171-3.319 (m, 2H), 3.620-3.701 (m, 1H),3.913 (d, J=7.1 Hz, 1H), 7.412-7.511 (m, 3H), 7.803-7.923 (m, 4H).¹³C-NMR (d₄-DMSO) analysis: δ 14.45, 20.31, 29.59, 42.57, 47.38, 76.67,125.77, 126.12, 126.19, 127.33 127.50, 127.62, 128.41, 132.31, 133.14,137.12 (Expected: 16C, Found: 16C). MS (CI⁺): Calcd for C₁₆H₂₂NOCl:279.14, Found: 244.2 (M-Cl).

Example 10 Resolution of 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamineinto the (+)-isomer

A 100 mL round bottom flask equipped with a stir bar was charged with1.52 g 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine (6.2 mmol), 0.979 g(S)-(+)-mandelic acid (6.4 mmol), and 30 mL ethanol. After stirring at60° C. for 1 hour, the homogenous mixture was allowed to cool to roomtemperature yielding a solid. The solid was collected by filtration andthen recrystallized twice from hot ethanol to yield thediastereomerically pure mandelate salt. The mandelate salt was treatedwith 2N NaOH to generate the free base, which was extracted three timeswith 50 mL diethyl ether. The extracts were pooled and treated with HClgas to yield the (+) isomer of3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine HCl salt (0.504 g, 28.9%).NMR and Mass spectral data were identical to that of3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. [α]=+30.3° (c 0.132,MeOH).

Example 11 Resolution of 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamineinto the (−)-isomer

A 100 mL round bottom flask equipped with a stir bar was charged with1.52 g 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine (6.2 mmol), 0.975 g(R)-(−)-mandelic acid (6.4 mmol), and 30 mL ethanol were combined asdescribed in Example 9 to yield the (−) isomer of3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine (0.493 g, 28.3%). NMR andMass spectral data were identical to that of3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine. [α]=−29.8° (c 0.134,MeOH).

Example 12 Synthesis ofN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, HCl salt

3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine (201 mg, 0.7 mmol) wasconverted to N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamineHCl salt (216 mg, 98% yield) using the N,N-dimethylation procedure ofCarlier et al. (Carlier et al., Bioorg. Med. Chem. Lett., 8:487-492(1998)) followed by treatment with methanolic HCl. ¹H-NMR (CD₃OD)analysis: δ 1.064-1.098 (m, 6H), 1.511-1.586 (m, 1H), 3.113 (s, 6H),3.538-3.683 (m, 2H), 4.062 (dd, J=7.2 Hz, 12.4 Hz, 1H), 4.130 (dd, J=1.9Hz, 9.3 Hz, 1H), 7.635-7.726 (m, 3H), 8.038-8.119 (m, 4H). ¹³C-NMR(CD₃OD) analysis: δ 14.80, 20.76, 31.46, 44.29, 44.85, 46.18, 62.28,80.70, 126.86, 127.65, 127.90, 128.93, 129.01, 129.08, 130.58, 134.72,135.43, 137.10 (Expected: 18C, Found: 18C). MS (CI⁺): Calcd forC₁₈H₂₆NOCl: 307.17, Found: 272.2 (M-Cl).

Example 13 Resolution ofN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine into the(+)-isomer

The (+) isomer of 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine (103 mg,0.37 mmol) was converted to the (+) isomer ofN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine HCl salt (114mg, 100%) using the N,N-dimethylation procedure of Carlier et al.(Carlier et al., Bioorg. Med. Chem. Lett., 8:487-492 (1998)) followed bytreatment with methanolic HCl. NMR and Mass spectral data were identicalto that of N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine.[α]=+4.2° (c 0.190, MeOH).

Example 14 Resolution ofN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine into the(−)-isomer

The (−) isomer of 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine (104 mg,0.37 mmol) was converted to the (−) isomer ofN,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine HCl salt (114mg, 100%) using the N,N-dimethylation procedure of Carlier et al.(Carlier et al., Bioorg. Med. Chem. Lett., 8:487-492 (1998)) followed bytreatment with methanolic HCl. NMR and Mass spectral data were identicalto that of N,N-dimethyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine.[α]=−3.8° (c 0.260, MeOH).

Example 15 Synthesis ofN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine, HCl saltSynthesis of Carbamate C

A weighed amount of 3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine istreated with ethyl chloroformate as described in Example 3 to yieldcarbamate C.

Reduction of carbamate C toN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine

A round bottom flask equipped with a stir bar and water condenser ischarged with a weighed amount of carbamate C, a weighed amount ofLiAlH₄, and a measured volume of dry THF. The mixture is refluxed underN₂ for several hours, and the reaction is quenched by adding water.After quenching the reaction, aqueous NaOH is added with stirring toyield a white solid and a clear organic layer. The organic layer isdecanted, and water is added to the solid. After mixing for 15 minutes,the white solid is extracted four times with diethyl ether, and theether extracts are pooled. The pooled extracts are dried over K₂CO₃,filtered, and concentrated in vacuo to yieldN-methyl-3-hydroxy-4-methyl-2-(2′-naphthyl)pentylamine free base. Thisfree base is converted to the HCl salt by treatment with concentratedHCl in methanol, followed by concentration in vacuo.

OTHER EMBODIMENTS

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

1. A composition comprisingN-methyl-3-hydroxy-2-(2′-naphthyl)-3-phenylpropylamine.